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COP and Clathrin-Coated Vesicle Budding: Different Pathways, Common Approaches Harvey T Mcmahon1 and Ian G Mills2

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COP and Clathrin-Coated Vesicle Budding: Different Pathways, Common Approaches Harvey T Mcmahon1 and Ian G Mills2 COP and clathrin-coated vesicle budding: different pathways, common approaches Harvey T McMahon1 and Ian G Mills2 Vesicle and tubule transport containers move proteins and of these coated vesicles have now been purified: COPI- lipids from one membrane system to another. Newly forming (coatomer) and COPII-coated vesicles (where COP transport containers frequently have electron-dense coats. stands for coat protein complex) and clathrin-coated Coats coordinate the accumulation of cargo and sculpt the vesicles [2–6] (Figure 1). Coat components are needed membrane. Recent advances have shown that components of for generation of highly curved membrane areas, recruit- both COP1 and clathrin-adaptor coats share the same structure ment of cargo (and exclusion of non-cargo proteins/ and the same motif-based cargo recognition and accessory lipids), vesicle scission and uncoating factor recruitment. factor recruitment mechanisms, which leads to insights on Not all budding pathways lead to small vesicles, but the conserved aspects of coat recruitment, polymerisation and high membrane curvature of these transporters makes membrane deformation. These themes point to the way in them intrinsically more fusogenic than larger vesicle which evolutionarily conserved features underpin these structures like macro-phagosomes. There may be many diverse pathways. ways to bud a membrane but we would argue that producing a small vesicle that concentrates cargo mole- Addresses cules requires a coat. 1MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK Just as a SNARE-based fusion underpins the hypothesis e-mail: [email protected] 2Department of Oncology, University of Cambridge, Hutchison MRC of a common mechanism for all membrane fusion, so a Cancer Research Centre, Hills Road, Cambridge, CB2 2XZ, UK coat-based budding hypothesis has emerged as a common e-mail: [email protected] theme in vesicle budding. This review highlights con- served mechanisms underpinning diverse budding path- ways and the differences that often reflect specificity and Current Opinion in Cell Biology 2004, 16:379–391 regulation. This review comes from a themed issue on Membranes and organelles Background to coated vesicle budding Edited by Judith Klumperman and Gillian Griffiths Clathrin-coated vesicles are named after the protein that Available online 20th June 2004 self-polymerises into a lattice around these vesicles as they bud from the plasma membrane, trans-Golgi net- 0955-0674/$ – see front matter work (TGN) and endosomes [7,8]. For all clathrin-coated ß 2004 Elsevier Ltd. All rights reserved. vesicles, clathrin is the central organiser (Box 1). It con- centrates cargo adaptors, leading to a diverse protein and DOI 10.1016/j.ceb.2004.06.009 lipid load in the forming vesicle, and its polymerisation into a curved lattice stabilises the nascent membrane bud Abbreviations AP adaptor protein as it forms. Each clathrin molecule contains an N- Arf ADP ribosylation factor substrate terminal domain that has a b-propeller structure and binds ARH autosomal recessive hypercholesterolemia protein to peptide motifs between its blades [9]. This multi- COP coat protein complex bladed propeller allows for multiple protein interactions ER endoplasmic reticulum with various specificities [10], recruiting a versatile array GGA Golgi-localised, g-ear containing, ADP-ribosylation factor-binding protein of different cargo adaptors and membrane attachment Hrs hepatocyte growth factor receptor tyrosine kinase substrate proteins [11,12,13 ,14]. TGN trans-Golgi network The main cargo adaptor proteins characterised to date are the classical adaptor protein (AP) complexes, AP1, AP2, Introduction: The coated vesicle budding AP3 and AP4 [15–19] (Table 1). Most AP complexes hypothesis adapt and/or link clathrin to selected membrane cargo and Early electron microscopy studies of cells led to a vesicle- lipids, and they also bind accessory proteins that regulate transport hypothesis for protein trafficking. Vesicular coat assembly and disassembly (such as AP180, epsins transport intermediates bud from a donor organelle and and auxilin; see Table 2). ‘Alternative adaptors’ can also then fuse with an acceptor organelle. Budding intermedi- function in clathrin-mediated vesicle budding. These ates were initially identified by their electron-dense adaptors are sometimes called ‘monomeric adaptors’ ‘coats’ and were found on the plasma membrane and but this does not accurately describe some of the mem- intracellular organelles [1] (Figure 1). Three major classes bers of this group, which are dimeric. GGA adaptors www.sciencedirect.com Current Opinion in Cell Biology 2004, 16:379–391 380 Membranes and organelles Figure 1 (a) (b) (d) (c) Current Opinion in Cell Biology Current Opinion in Cell Biology 2004, 16:379–391 www.sciencedirect.com COP and clathrin-coated vesicle budding McMahon and Mills 381 Box 1 Definitions factors (Figure 1 and Table 1) [20,21]. GGAs and Hrs (hepatocyte growth factor receptor tyrosine kinase sub- Clathrin Clathrin is a trimer with a central hub domain from which extends strate) have similar but not identical domain compositions three legs ending in terminal (b-propeller/WD40) domains. This (Figure 1). By analogy with cargo binding to the GGA building block of a cage structure is known as a triskelion. During VHS domain, Hrs may also bind cargo, but this protein is polymerisation the legs on neighbouring triskelia twist around each targeted via its FYVE domain to endosomes. Arrestins, other and make a lattice. Recruitment proteins bind to these terminal domains and help to concentrate clathrin on membranes. epsins, disabled-2 and ARH (autosomal recessive Concentrated clathrin self-assembles [7]. hypercholesterolemia protein) can also be classified as alternative adaptors as they also link cargo and mem- Cargo adaptors Proteins that link cargo into the clathrin-coated pit branes to the clathrin lattice (Table 2) [11,13 ,14,22]. These may work in some cases alongside or indepen- Classical clathrin adaptors: AP1, AP2, AP3 and AP4 complexes each have four subunits: two dently of classical adaptors to recruit their cargo into large, one medium and one small, which are believed to be stably budding vesicles [23 ]. Since multiple adaptors are found associated in the cell. AP1 is involved in protein sorting from the TGN in individual coated pits, it is not necessary for all adaptors and endosomes; AP2 traffics from the plasma membrane; AP3 to have a direct clathrin interaction. traffics to lysosomes and AP4 is found on vesicles in the vicinity of the TGN [19]. COP coats come in two very different flavours, belying Alternative clathrin adaptors the homologous nomenclature (Figure 1 and Table 1). Monomeric and dimeric cargo/clathrin adaptors like GGAs and Hrs COPI coat components have sequence homology to cla- Accessory proteins thrin AP complex proteins [24–28]. Just as clathrin and Clathrin recruitment, membrane bending and scission molecules. This group can cover all proteins involved in clathrin-mediated adaptors come together to form clathrin-coated vesicles, endocytosis except clathrin. But the definition is more usefully so the two subcomplexes of COPI coats come together to applied when it neither covers clathrin nor cargo adaptors. recruit cargo into buds [28]. COPI coat components traffic COP coat subcomplexes primarily from the Golgi to the endoplasmic reticulum There are seven COPI subunits (a, b, b’, d, g, e and z) divided into two (ER) and between Golgi cisternae. subcomplexes: the F subcomplex composed of b, d, g and z-COPs, of which the b and g subunits display homology to AP complex COPII-coated vesicles traffic from the ER to the Golgi. appendage domains, and the B subcomplex, within which the a and b’ subunits contain WD40 repeats akin to clathrin terminal There are again two subcomplexes (Figure 1 and Table 1) domains [28]. that come together to recruit cargo into buds. In the clathrin-like subcomplex of COPI coats (the B-subcom- COPII coat proteins can also be considered as subcomplexes consisting of Sec13/31p and Sec23/Sec24p. Both Sec13/31p have plex) and in the Sec13/31p subcomplex of COPII coats WD40 repeats akin to clathrin whereas Sec23/24p functionally there are b-propeller domains (made from WD40 repeats) resemble the AP complexes in a cargo recruitment capacity. [29] that form a protein interaction face just like the b- Motif domains propeller clathrin terminal domains. COP coats are there- Generally regions lacking tertiary structure in proteins containing fore also multisubunit protein complexes, whose compo- short sequence motifs that serve as binding sites for ligand. This is a nents select cargo, drive membrane curvature and vesicle generic term that can be applied to all proteins with similar domains budding (Table 1). We will now look at some common- across the proteome and stands in contrast to ‘structured domains’. It is likely that many of these domains have a limited structure. alities in these diverse vesicle budding pathways. Conservation of coat recruitment and (Golgi-localized, g-ear containing, ADP-ribosylation- membrane curving mechanisms factor-binding proteins) are associated with TGN clathrin A long-recognised commonality between COPI, COPII coats and bind cargo, membranes, clathrin and accessory and many clathrin budding pathways is that small (Figure 1 Legend) Structural and functional homology between COP1 coats and clathrin-adaptor coats. (a) The ‘classical’ AP2 clathrin adaptor with its four subunits forms a complex that links cargo recruitment to clathrin. Clathrin is a trimer and each terminal domain is represented as being bound to a b2 adaptin appendage and hinge domain. The other heterotetrameric AP complexes, AP1, AP3 and AP4, have the same overall structure. (b) The COPI F-subcomplex probably has a similar topology to the AP complex and the B-subcomplex is likely to be the functional equivalent of clathrin. (c) Likewise, COPII has two subcomplexes, Sec23/24 and Sec13/31. (d) Alternative clathrin adaptors GGAs and Hrs are like the large subunits of AP complexes in that they bind to cargo, clathrin and membranes.
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